BACKGROUND: In recent years, due to the side effects of synthetic blood pressure-lowering drugs and synthetic antioxidants, many studies have been conducted on their natural substitutes. Milk proteins contain peptide components called bioactive peptides that can play a significant role in inhibiting oxidation and lowering blood pressure. OBJECTIVES: In this study, milk was hydrolyzed using a broccoli protease extract to produce bioactive peptides with angiotensin converting enzyme inhibitory (ACE-I) and antioxidant effects. METHODS: Broccoli protease extract was added to defatted milk at concentrations of 0.5, 1, and 2 U per 100 mL and the degree of hydrolysis was determined using the OPA method. Antioxidant activity was evaluated by the DPPH radical scavenging assay, and angiotensin converting enzyme inhibitory (ACE I) activity was determined using the HHL substrate method. Physicochemical and sensory properties of milk were also assessed. RESULTS: The results showed that treatment of milk with broccoli protease extract significantly increased the degree of hydrolysis and led to the formation of bioactive peptides with antioxidant and ACE inhibitory activities. The highest ACE inhibitory activity (35%) and DPPH radical scavenging activity (78%) were observed in the milk treated with 2 U enzyme. CONCLUSION: These findings suggest that broccoli-derived protease may serve as a promising natural enzyme for the production of bioactive peptides in dairy systems.